Fabrication of biomimetic placental barrier structures within a microfluidic device utilizing two-photon polymerization

Abstract

The placenta is a transient organ, essential for development and survival of the unborn fetus. It interfaces the body of the pregnant woman with the unborn child and secures transport of endogenous and exogenous substances. Maternal and fetal blood are thereby separated at any time, by the so-called placental barrier. Current in vitro approaches fail to model this multifaceted structure, therefore research in the field of placental biology is particularly challenging. The present study aimed at establishing a novel model, simulating placental transport and its implications on development, in a versatile but reproducible way. The basal membrane was replicated using a gelatin-based material, closely mimicking the composition and properties of the natural extracellular matrix. The microstructure was produced by using a high-resolution 3D printing method – the two-photon polymerization (2PP). In order to structure gelatin by 2PP, its primary amines and carboxylic acids are modified with methacrylamides and methacrylates (GelMOD-AEMA), respectively. High-resolution structures in the range of a few micrometers were produced within the intersection of a customized microfluidic device, separating the x-shaped chamber into two isolated cell culture compartments. Human umbilical-vein endothelial cells (HUVEC) seeded on one side of this membrane simulate the fetal compartment while human choriocarcinoma cells, isolated from placental tissue (BeWo B30) mimic the maternal syncytium. This barrier model in combination with native flow profiles can be used to mimic the microenvironment of the placenta, investigating different pharmaceutical, clinical and biological scenarios. As proof-of-principle, this bioengineered placental barrier was used for the investigation of transcellular transport processes. While high molecular weight substances did not permeate, smaller molecules in the size of glucose were able to diffuse through the barrier in a time-depended manner. We envision to apply this bioengineered placental barrier for pathophysiological research, where altered nutrient transport is associated with health risks for the fetus.

Impact of Hydrogel Stiffness on Differentiation of Human Adipose-Derived Stem Cell Microspheroids

“…The methacrylation was carried out as already described. 33 Samples were exposed to 25 mW/cm 2 UV-A light (LITE-Box G136 365 nm; NK-OPTIK, Germany) for 10 min to induce hydrogel cross-linking. Afterwards 0.5 mL of control medium per gel clot was added and the dishes were transferred to the incubator.…”

Advances on Bone Substitutes through 3D Bioprinting

“…This method enables the production of 3D structures in microfluidic devices, well resembling the native microenvironment [ 184 , 185 ].…”

Methacrylation increase growth and differentiation of primary human osteoblasts for gelatin hydrogels

“…L-Lysine monohydrochloride was purchased from Merck (Darmstadt, Germany), dithiothreitol from Thermo Fisher Scientific (Waltham, USA), sodium azide (99%), and iodine (I2) from Avocado Research Chemicals Ltd. (Karlsruhe, Germany) were prepared as reported [18,23,24]. The methacrylation was carried out as already described [25]. Samples were exposed to a 25-mW/cm 2 UV-A light (LITE-Box G136 365 nm; NK-OPTIK, Germany) for 10 min to induce hydrogel crosslinking.…”